LOCUS       V004067                 8600 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004067
VERSION     V004067
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8600)
  AUTHORS   Kyriakakis P, Catanho M, Hoffner N, Thavarajah W, Jian-Yu V, Chao
            SS, Hsu A, Pham V, Naghavian L, Dozier LE, Patrick G, Coleman T.
  TITLE     Biosynthesis of Orthogonal Molecules Using Ferredoxin and
            Ferredoxin-NADP+ Reductase Systems Enables Genetically Encoded PhyB
            Optogenetics
  JOURNAL   ACS Synth Biol (2018) In press
   PUBMED   29301067
REFERENCE   2  (bases 1 to 8600)
  AUTHORS   Kyriakakis P.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-NOV-2017) Bioengineering, UCSD, 9500 Gilman drive,
            PFBH room 251 MC0412, La Jolla, CA 92093, United States of America
REFERENCE   3  (bases 1 to 8600)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8600)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
            Biol (2018) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (15-NOV-2017) Bioengineering, UCSD, 9500 Gilman drive, PFBH room 251
            MC0412, La Jolla, CA 92093, United States of America"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        655..673
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             732..881
                     /label="VP64"
                     /note="tetrameric repeat of the minimal activation domain
                     of herpes simplex virus VP16 (Beerli et al., 1998)"
     misc_feature    888..905
                     /label="linker 1"
                     /note="linker 1"
     CDS             906..926
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             1356..1712
                     /label="RelA (p65) AD"
                     /note="transcriptional activation domain of human RelA,
                     also known as p65 (O'Shea and Perkins, 2008)"
     misc_feature    1713..1730
                     /label="linker 2"
                     /note="linker 2"
     CDS             1731..2300
                     /label="Rta AD"
                     /note="transcriptional activation domain from the human
                     herpesvirus 4 (Epstein-Barr virus) replication and
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992;
                     Chavez et al., 2015)"
     CDS             2340..3908
                     /gene="PIF3"
                     /label="Transcription factor PIF3"
                     /note="Transcription factor PIF3 from Arabidopsis thaliana.
                     Accession#: O80536"
     promoter        complement(3944..3962)
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3988..4212
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4258..4686
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4700..5030
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             5097..5888
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    6065..6198
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6235..6251)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6259..6275)
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6283..6313)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6328..6349)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6637..7225)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7399..8256)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8257..8361)
                     /label="AmpR promoter"
ORIGIN
        1 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg actatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
      661 gactcactat agggagaccc aagcttggta ccgagctcgg atccatggag gctagtggat
      721 ctgggagagc agatgctctg gatgactttg atctggacat gctgggctcc gacgcactcg
      781 atgacttcga tctggatatg cttggcagcg acgcactgga cgacttcgat ctggacatgt
      841 tgggatctga cgcgctggac gacttcgatc tggacatgct gatcaattct cgctcatccg
      901 gttcaccaaa gaaaaagagg aaggttggga gtcagtatct tcccgatacc gacgatcggc
      961 accggataga ggaaaagagg aagcggacat acgagacctt caaatctatc atgaagaagt
     1021 cacccttctc tggacccacc gatcccagac ccccgccccg gcgcattgcc gtaccctccc
     1081 gcagtagcgc ctcagtcccc aagcctgcac ctcaacctta tccttttacc agttcactgt
     1141 ctaccattaa ttacgacgag tttccgacga tggtgtttcc ctctggccag atatctcagg
     1201 cctctgccct ggcgcctgca cctccacagg tactgcctca ggcaccggca cccgccccgg
     1261 ctcctgcaat ggtgtccgcc ctcgctcagg cccccgcacc tgtgcctgta cttgcccctg
     1321 gccccccgca ggctgtcgct cctcctgccc ccaagcctac ccaggccggg gagggaactc
     1381 tgtcagaggc cttgttgcag ttgcagttcg atgacgagga cctgggagct cttctgggca
     1441 attctaccga ccccgccgtc ttcactgacc tggcaagcgt ggacaatagc gaatttcaac
     1501 agttgcttaa ccagggtatt ccggttgctc cgcatactac tgagcccatg ctgatggaat
     1561 accctgaggc catcaccaga ttggtcaccg gggcacagcg gcctccggac ccagctcccg
     1621 cccctctggg cgcccccgga ctgccaaacg gcctgctgtc cggagacgaa gatttctcta
     1681 gtattgccga catggacttt tctgcccttt tgggcagcgg gagcggttcc agagatagtc
     1741 gcgaaggaat gttcctgcca aaacccgaag caggcagcgc cataagtgac gtgttcgagg
     1801 gacgggaagt gtgccagcca aagaggatta gaccattcca cccaccaggc tctccctggg
     1861 ctaataggcc cctccctgct tcattggcgc ctactccgac tggtcccgtc cacgagccag
     1921 tggggtctct gactcctgca cctgtgccac agcccctgga cccagcccca gctgtcaccc
     1981 ccgaggccag ccacctgctc gaagatccag atgaggaaac ctcccaggcc gtaaaggcct
     2041 tgcgggaaat ggccgatact gtaatcccac agaaagagga ggccgccatt tgtgggcaaa
     2101 tggatctcag tcatccccct ccaaggggtc acctggatga attgaccaca accttggaat
     2161 caatgacaga agacctcaat ctggatagcc ccctgacgcc ggagctgaac gagattcttg
     2221 acacttttct caacgatgaa tgcctcttgc atgccatgca tatcagcacc ggcctgtcta
     2281 tttttgatac ctccttgttt gacagcgccg gcagcgccgg cagcgccggc agcgaattca
     2341 tgcctctgtt tgagcttttc aggctcacca aagctaagct tgaatctgct caagacagga
     2401 acccttctcc acctgtagat gaagttgtgg agctggtgtg ggaaaatggt cagatatcaa
     2461 ctcaaagtca gtcaagtaga tcgaggaaca ttcctccacc acaagcaaac tcttctagag
     2521 ctagagagat tggaaatggc tcaaagacga ctatggtgga cgagatccct atgtcagtgc
     2581 catcactaat gacgggtttg agtcaagacg atgactttgt tccatggttg aatcatcatc
     2641 cctcccttga tggatattgc tctgatttct tgcgtgatgt gtcgtctcct gttactgtca
     2701 acgagcaaga gagtgatatg gcggtaaacc aaactgcttt cccgttgttt cagagaagaa
     2761 aggatggcaa tgaatcagct cctgctgctt cttcgtcgca gtataacggt ttccaatcgc
     2821 attctctgta tggaagtgat agagctagag atcttcctag ccaacaaacc aatccggatc
     2881 ggtttactca gacgcaggaa ccactaatta ctagtaacaa gcctagtttg gtcaactttt
     2941 cacatttctt acgccctgca acttttgcga agactactaa taataacctt catgacacta
     3001 aagaaaagag tcctcaaagc ccgccaaatg tgtttcagac cagagttctt ggagctaaag
     3061 actctgaaga taaggttctt aacgagtctg ttgcttctgc tacgcctaaa gataaccaaa
     3121 aggcttgcct aatatcagag gactcatgta gaaaagacca agagagtgaa aaagcagttg
     3181 tatgttcttc tgttggctcg ggtaatagtc tcgatggccc atccgaaagt ccttcacttt
     3241 ctttaaagag aaagcattcg aatattcaag acattgactg tcatagtgaa gatgtggaag
     3301 aagaatcagg agatggaaga aaggaagcag gtccatctcg aacgggtttg ggttcaaaga
     3361 gaagccgctc tgcagaagtg cataatctgt ctgaaaggag acggcgtgat aggatcaacg
     3421 agaagatgcg tgccctgcaa gaactcattc caaactgtaa caaggtggac aaagcttcga
     3481 tgctagatga agccatcgag tatctcaagt cactccaact tcaagtgcag atcatgtcaa
     3541 tggcgtctgg ttactatctg ccaccggcgg ttatgttccc accgggtatg gggcattacc
     3601 cggcagcagc tgctgcaatg gcaatgggta tgggaatgcc ttatgcaatg ggcttgcctg
     3661 atttgagccg tggtggttca tcggttaacc acggaccaca gttccaagtc tcggggatgc
     3721 aacaacaacc agtggcgatg ggtattccac gtgtctctgg tggtggtatc tttgccggtt
     3781 cttcgacgat tggcaatggc tcgactagag atttatctgg ttctaaagat caaacaacga
     3841 cgaataacaa cagtaacttg aaaccaataa agagaaaaca ggggtcttct gatcagtttt
     3901 gtggatcgtg cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa
     3961 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     4021 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     4081 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     4141 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     4201 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     4261 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4321 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4381 tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg
     4441 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4501 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4561 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4621 ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4681 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg
     4741 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4801 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4861 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4921 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4981 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     5041 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     5101 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5161 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5221 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     5281 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5341 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5401 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5461 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5521 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5581 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     5641 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5701 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5761 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5821 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5881 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5941 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6001 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6061 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6121 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6181 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6241 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6301 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6361 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6421 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6481 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6541 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6601 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6661 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6721 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6781 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     6841 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6901 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6961 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7021 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7081 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7141 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7201 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7261 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7321 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7381 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7441 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7501 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7561 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7621 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7681 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7741 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7801 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7861 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7921 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7981 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8041 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8101 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8161 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8221 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8281 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8341 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     8401 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     8461 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     8521 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     8581 aggcgttttg cgctgcttcg
//
